PTMScan ® Ubiquitin Remnant Motif (K-epsilon-GG) Kit

Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase, solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan® Motif antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan® IAP Buffer #9993 included in the kit.

PTMScan® Ubiquitin Remnant Motif (K-epsilon-GG) Kit has a higher sensitivity and specificity magnetic bead version: PTMScan® HS Ubiquitin/SUMO Remnant Motif (K-epsilon-GG) Kit in 10-assay (#59322) or 3-assay (#19089) formats, as well as a bead-only (#34608) format.